Insights into Metabolites Profiling and Pharmacological Investigation of Aconitum heterophyllum wall ex. Royle Stem through Experimental and Bioinformatics Techniques.

The Aconitum genus is a leading source of a wide range of structurally diverse metabolites with significant pharmacological implications. The present study investigated metabolite profiling, pharmacological investigation, anticancer potential, and molecular docking analysis of the stem part of Aconitum heterophyllum (AHS). The metabolite profiling of the AHS extract was experimentally examined using LC-MS/MS-orbitrap in both modes (ESI+/ESI-) and GC-MS in EI mode. The in vitro MTT model was used to study the anticancer potential, while the in vivo animal model was used to study the anti-inflammatory and antinociceptive activities. The MOE software was used for the molecular docking study. A total of 118 novel and previously known metabolites, among 44 metabolites (26 in ESI+ positive mode and 18 in ESI- negative mode) in the MeOH extract, while 74 metabolites (46 in ESI+ and 28 in ESI- mode) were identified in the n-hexane extract via LCMS/MS. The identified metabolites include 24 phenolic compounds, 18 alkaloids, 10 flavonoids, 24 terpenoids, 2 coumarins, 2 lignans, and 38 other fatty acids and organic compounds. The major bioactive metabolites identified were hordenine, hernagine, formononetin, chrysin, N-methylhernagine, guineesine, shogaol, kauralexin, colneleate, zerumbone, medicarpin, boldine, miraxinthin-v, and lariciresinol-4-O-glucoside. Furthermore, the GC-MS study helped in the identification of volatile and nonvolatile chemical constituents based on the mass spectrum and retention indices. The methanol extract significantly inhibited tumor progression in H9c2 and MDCK cancer cells with IC50 values of 186.39 and 199.63 µg/mL. In comparison, the positive control aconitine exhibited potent IC50 values (132.32 and 141.58 µg/mL) against H9c2 and MDCK cell lines. The anti-inflammatory (carrageenan-induced hind paw edema) and antinociceptive (acetic acid-induced writhing) effects were significantly dose-dependent, (p < 0.001) and (p < 0.05), respectively. In addition, a molecular docking study was conducted on identified ligands against the anti-inflammatory enzyme (COX-2) (PDB ID: 5JVZ) and the cancer enzyme ADAM10 (PDB ID: 6BDZ) which confirmed the anti-inflammatory and anticancer effects in an in silico model. Among all ligands, L2, L3, and L7 exhibit the most potent potential for inhibiting COX-2 inflammation with binding energies of -7.3424, -7.0427, and -8.3562 kcal/mol. Conversely, against ADAM10 cancer protein, ligands L1, L4, L6, and L7, with binding energies of -8.0650, -7.7276, -7.0454, and -7.2080 kcal/mol, demonstrated notable effectiveness. Overall, the identified metabolites revealed in this AHS research study hold promise for discovering novel possibilities in the disciplines of chemotaxonomy and pharmacology.

Evaluation of Acacia cyanophylla for their analgesic, anti-pyretic and anti-inflammatory potentials.

The present paper presents results of analgesic, antipyretic activity and anti-inflammatory potential of extract obtained from Acacia cyanophylla when tested at different doses. Analgesic potential of the crude methanolic extract tested by acetic acid assay was dose dependent and maximum activity of 61.60% was measured at 400 mg/kg. Analgesic activity by hot plate method revealed that maximum activity of 36.98% was noted when the mice were exposed to 90 minutes at higher dose of 400 mg/kg. Similar pattern for antipyretic activity was observed as noted for analgesic activity. Anti-inflammatory activity was dose and time dependent when evaluated by Carrageenan-induced paw edema and Xylene-induced ear edema model. Maximum anti-inflammatory activity (43.32%) was shown by crude methanolic extract of Acacia cyanophylla at 400mg/kg-1 after 5 hours on Carrageenan-induced paw edema model. Similarly, maximum (68.80%) anti-inflammatory activity was noted when accessed by Xylene-induced ear edema model at the dose of 200mg/kg after 60 minutes. No in vivo toxicity of the extracts up to the dose of 2000mg/kg was observed using albino mice.

Antimicrobial efficacy and prevalence of colicinogenic E. coli in faecal matter of human, cow and sheep.

Colicinogenic Escherichia coli in the human or animal gut prevent pathogen accumulation, thereby protecting against severe gut infections. This ability of commensal human and animal E. coli to restrain Gram-negative pathogens was evaluated invitro. Approximately 13.2% of E. coli isolates inhibited the growth of target pathogens. The greatest inhibition of 36% was observed against E. coli O157:H7 followed by 35%, 27%, 24% and 13% inhibition against E. coli O26:H11, Pseudomonas aeruginosa, Salmonella enterica and Klebsiella pneumoniae, respectively. Detection of 20 colicin determinants in colicinogenic E. coli revealed that >50% isolates exhibited multiple colicin genes. Among all isolates, 70.4% of E. coli had the col E6 gene followed by col Ib (66.4%), E4 (53.6%), E7 (49.9%), J (35.3%) and M (35.2%) determinants. The frequency of col D (8.8%), Ia (27.9%), S4 (12%), E3 (13.2%) and E9 (2.9%) was greater in human samples compared with samples from cows and sheep, whereas col10 (5.8%) and E5 (4.4%) were produced only by cow-derived E. coli. Colicinogenic E. coli belonging to phylogenetic group B2 (52.8%) were more prevalent followed by D1 (16%), B1 (13.2%), A1 (11.6%) and A0 (5.8%). The 16S rRNA sequencing of all colicinogenic E. coli provided 27 non-clinical variants of E. coli that can be further explored for their probiotic properties to minimize risk of gut diseases.

Antibacterial activity of the crude extracts from medicinally important Thuja occidentalis.

The present research was carried out at the Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, KPK Pakistan. In this study crude methanolic extracts from Thuja occidentalis were tested for their antimicrobial activity against five different bacterial strains (Bacillus subtilis, Escherichia coli, Klebsiella pneumonia, Xanthomonas sp. and Staphylococcus aureus) at two different concentrations (10 and 20 mg/ml) using disc diffusion assay. The results showed that Klebsiella pneumoniae was most sensitive to crude extracted sample from leaves at both low and high concentrations measuring 37 and 57% zone of inhibition respectively and Xanthomonas and Staphylococcus aureus was found to be more resistant to the crude extracted samples from leaves at both concentrations. Similarly, Klebsiella pneumoniae was most sensitive to crude extracted samples from seeds at high concentration followed by E. coli at both concentrations. Staphylococcus aureus on the other hand was found to be more resistant to the crude extracted samples at low and high concentrations. These results suggested that extracts prepared from the leaves and seeds of Thuja occidentalis can be used as natural remedy for the treatment of various bacterial infections.

Antimicrobial activity of different solvent extracted samples from the leaves and fruits of Capsicum annuum.

The current research describes the antimicrobial potential of methanol, n-hexane, n-butanol, ethyl acetate and aqueous extracted samples from the leaves and fruits tissues of Capsicum annuum. Different solvent extracted samples were screened against six pathogenic microorganisms including five bacterial and one fungal specie by disc diffusion susceptibility assay using 1, 2 and 3 mg disc-1 concentrations. When analyzed statistically the data showed that different solvent extracted samples from both leaves and fruits of Capsicum annuum revealed varying degrees of antibacterial and antifungal activities. n-butanol and ethyl acetate extracted fractions from both leaves and fruits showed significant inhibition of growth against all the tested microorganisms at 1, 2 and 3 mg disc-1 concentrations. Escherichia coli were completely resistant to aqueous extracts obtained from the leaves at all the three concentrations. Klebsiella pneumonia was resistant to n-hexane extracted fraction from leaves at 1mg disc-1 concentration, however, was susceptible at 2 and 3 mg disc-1 concentrations. The growth of Pseudomonas aeruginosa and Staphylococcus aureus were effectively inhibited by all the solvent extracted fractions from the fruits while aqueous fraction was not able to inhibit the growth of Bacillus subtilis. The growth of Candida albicans was effectively inhibited by ethyl acetate extracted fraction from leaves at 3 mg disc-1 concentration.

Anti-inflammatory, anti-pyretic and analgesic activities of Tamarix dioica.

Medicinal and aromatic plants contribute to major portion of the flora. The plant materials obtained from these plants are used in the pharmaceuticals, cosmetics, and drug industries. Tamarix dioica is locally used in the management of splenic and hepatic inflammation as well as diuretic and carminative. It also possesses cytotoxic, antimicrobial, antifungal and anti-inflammatory activity. The present study investigates the anti-inflammatory, anti-pyretic and analgesic activities of the crude methanolic extract from Tamarix dioica. Anti-inflammatory activity was measured by Carrageenan Induced Paw Edema and Xylene Induced Ear Edema methods. Pyrexia induction with Brewer's yeast assay was used to determine antipyretic activity and analgesic activity was estimated by acetic acid induced writhing and hot plate methods. The data indicated that anti-inflammatory, anti-pyretic and analgesic activities of the crude methanolic extract from Tamarix dioica was dose and time dependent when measured by different assays. Exposure of model animal to increasing concentrations of the plant extract for longer period increased their anti-inflammatory, anti-pyretic and analgesic activities. Significantly highest anti-inflammatory, anti-pyretic and analgesic activities were noted at highest doses of the crude methanolic extract for longer exposure compared with their respective controls.

GC-MS analysis of bioactive compounds present in medicinally important Periploca hydaspidis.

The present study investigates the pharmaceutically bioactive compounds in Methanol, n-hexane and ethyl acetate extracted samples from the root of Periploca hydaspidis through Gas Chromatography and Mass Spectroscopy analysis. The mass spectrum obtained was compared with the data base of National Institute of Standards and Technology (NIST) which contains more than 62000 patterns of the mass spectrum. During matching with NIST library the match factor greater than 700 was considered only for better and pure results. The GC-MS analysis revealed the presence of various important compounds in the extracts like Lupeol, Furanol, Decanal, Decanoic acid, Dioxane and Oxirane. Besides these compounds the analysis also revealed the presence of antibiotics, fatty acids and protein.

Bioinspired synthesis and characterization of gold nano-particles from medicinally important Periploca hydaspidis and their in vitro antioxidant and antimicrobial activity.

This research investigates the synthesis and characterization of gold nanoparticles from Periploca hydaspidis and their antimicrobial and anti oxidant activity. The synthesis of AuNPs was confirmed by UV-Vis spectrophotometer and structure by a high resolution atomic force microscope. X-ray diffraction and Fourier transformed infrared spectroscopy was used to study the crystallite size and different functional groups. DPPH radical scavenging activity and disc diffusion protocol was applied for the determination of antioxidant and antimicrobial activity. A ratio of 1:8 of 1mM AuCl3 solutions with plant boiled extract used for synthesis of gold nano-particles. The formation of the gold nano-particles was determined by the color change from yellow to dark purple which were confirmed by UV-Vis spectrophotometer. Gold nano-particles were stable between 24°C and 39°C, mM concentration of the salt and neutral pH. The groups responsible for the synthesis of gold nano-paricles were Alkenes and aliphatic amines. The AuNP were cubic in nature and the nanocrystallite size was 6.99nm. Gold nano-particles revealed good antioxidant activity and controlled the growth of K. pnemoniae, E. coli, X. compestris, C. albicans and P. chrysogenum.

In vitro antimicrobial activity and phytochemical analysis of different solvent extracted samples from medicinally important Litsea glutinosa.

The present research work investigate the antimicrobial activities of crude methanolic extracted samples from the leaves of Litsea glutinosa against different microorganisms through disc diffusion assay applied in two different concentrations of 1 and 2mg disc1. The tested microbial species included B. subtilis, E. coli, P. aeruginosa, K. pneumoniae and C. albicans. The crude methanolic extract was applied in two different concentrations of 1 and 2mg disc1. Analysis of the data revealed that crude methanolic extracted samples showed different ranges of antimicrobial activities against all the tested microbes at both concentrations. Maximum growth inhibition was measured against gram negative Pseudomonas aeruginosa followed by the fungal specie Candida albicans. In case of Petroleum extracted fractions maximum growth reduction was measured in Candida albicans at higher concentration. Similarly, growth inhibition was more in Pseudomonas aeruginosa at higher concentration of aqueous extracted samples. Different solvent extracted samples showed the presence of alkaloids, flavonoids, proteins, fats, oils, tannins, carbohydrates, sterols and saponins.

Huperzine-A response to cognitive impairment and task switching deficits in patients with Alzheimer's disease.

BACKGROUND: Alzheimer's Disease (AD) is associated with cognitive decline due to various pathological mechanisms. There are several acetylcholinesterase inhibitor compounds which can improve cognition, but Huperzine-A is a natural sesquiterpene alkaloid extracted from Chinese herb (Huperzia Serrata) which has rapid action. METHODS: Double blind study was conducted. Participants included 50 patients with AD and 50 healthy individuals. Patients were recruited from Civil and BV hospital Bahawalpur and Nishter hospital Multan, Pakistan during May 2017 until February 2018 who were stable on Huperzine-A medication. Patients were tested twice. First, at the time of diagnosis to determine baseline scores. Second, post eight weeks of Huperzine-A treatment. Healthy individuals had single testing session. Participants completed Addenbrooke's Cognitive Examination and Trail Making Test. RESULTS: Patients with AD showed cognitive and task switching deficits in contrast with healthy individuals. There was significant improvement in cognition and task switching abilities post Huperzine-A treatment compared with baseline performance. CONCLUSION: Huperzine-A is effective in reducing cognitive and task switching deficits in patients with AD.

Effect of different solvent extracted samples from the leaves and fruits of Datura stramonium on the growth of bacteria and fungi.

Different solvent extracted samples from the leaves and fruits tissues of D. stramonium were tested against five pathogenic microorganisms by disc diffusion susceptibility method using 1, 2 and 3mg disc-1 concentrations. Methanol and chloroform extracted fractions from both leaves and fruits measured good growth inhibition of all the tested microorganisms at all concentrations. Bacillus subtilis was very resistant to n-butanol and aqueous extracted fractions of fruits tissues at all the tested three concentrations. The growths of Pseudomonas aeruginosa and Klebsiella pnuemonia were efficiently reduced by all the solvent extracted fractions from the fruits while aqueous fraction was unable to restrain the growth of Bacillus subtilis. The growth of Candida albicans was effectively reduced by aqueous extracted fraction from the leaves tissue at the highest concentration. Maximum growth reduction of (77%) was shown by chloroform extracted fractions from the leaves against Klebsiella pneumonia at 3mg disc-1 concentration. Minimum zone of inhibition (35.4%) was measured by n-butanol extracted fractions from the leaves against Pseudomonas aeruginosa at the lowest concentrations of 1mg disc-1. In case of leaves the most vulnerable bacteria was Bacillus subtilis while in case of fruits Klebsiella pneumonia was the most susceptible while Bacillus subtilis was the most resistant one.

Extraction and isolation of important bioactive compounds from the fruit of Physalis ixocarpa.

The current study investigates pharmaceutically important bioactive compounds in the fruits of Physalis ixocarpa. Two different extractions methods were used to study its effect on percent extract yield, recovery of bioactive compounds and antioxidant activity of the extracts. The data indicated that Soxhlet extraction had high efficiency of recovery than maceration method for extracting compounds; percent extract yield and antioxidant activity of the extracts. In maceration, the percent extract yield was found to be in order of water >methanol >ethyl acetate whereas in Soxhlet extraction, it was in order of methanol >water >ethyl acetate. Ethyl acetate extract produced by Soxhlet extraction showed strong antioxidant activity of 59.7% (250ppm) as compared to other extracts. Analysis of ethyl acetate extract showed the presence of Triglyceride. GC-MS study of triglyceride revealed the presence of trilinoleinic acid (9,12-Octadecdienoic acid), tripalmitin (hexadecanoic acid) and trioleinic acid (9-Octa decenoic acid). Four impure and three pure compounds were isolated from crude methanol extract of the fruit. The structure of pure compounds were identified by NMR and characterized as sugar, glucose and fructose.

In vitro antibacterial activity and phyto-chemistry of samples from the roots of Viola pilosa.

Anti-microbial activity and phytochemical analysis of samples from the roots of Viola pilosa was studied against six strains of bacteria. Data indicated that the tested bacterial strain differed in its sensitivity to the root extracts of Viola pilosa. The results showed that Xanthomonas campestris, Bacillus subtilis and S. aureus were more susceptible to butanol extracted fraction. S. aureus was totally resistant to aqueous extracted fraction at all concentrations and .Xanthomonas campestris and Bacillus subtilis were least susceptible to the same fraction at 0.5 mg disc-1. Similarly, ethyl acetate at 2 mg disc-1 concentrations was effective against Pseudomonas aeruginosa and Escherichia coli. Klebsiella pneumoniae was completely resistant to all the tested concentrations. Phytochemical analysis of the different solvent extracted samples suggested the presence or absence of different various metabolites including alkaloids, saponins, tannins, sterols, flavonoids, protein, carbohydrates and fats.

Impact of different solvent extracts from leaves and fruits of Eucalyptus globulus on growth of different bacteria and fungi.

The present research investigates the antimicrobial activities of the samples extracted from the leaves and fruits of Eucalyptus globulus through disc diffusion susceptibility assay using 1, 2 and 3mg disc-1 concentrations. Different extracted samples from the leaves and fruits of Eucalyptus globulus exhibited different degrees of antimicrobial. The data indicated that n-butanol and ethyl acetate extracted fraction of both the leaves and fruits inhibited the growth of all microorganisms at all the tested concentrations. Aqueous extracted sample of the leaves inhibited the growth of Candida albicans while the same fraction from the fruits showed no activity against Bacillus subtilis at any concentration. N-hexane extracted samples of the leaves inhibited the growth of Bacillus subtilis, E. coli and Pseudomonas aeruginosa at the tested concentrations while no activity was recorded against Klebsiella pneumonia, Candida albicans and Stephylococcus aureus. N-butanol extracted samples from the leaves and fruits showed activity against Pseudomonas aeruginosa at the tested concentrations. In case of leaves, the most susceptible bacteria was Bacillus subtilis (gram positive) and Stephylococcus aureus (gram positive) was the most resistant one. In case of fruits the most susceptible bacteria was Stephylococcus aureus (gram positive) and E. coli (Gram negative) was the most resistant one.

GC-MS profile of bioactive compounds from medicinally important Periploca hydaspidis.

The aim of the present study was to investigate bioactive compounds in different solvent extracted samples from the stem tissues of P. hydaspidis using Gas Chromatography and Mass Spectroscopy techniques. GC-mass spectrum was compared with the data base of National Institute of Standards and Technology (NIST) containing more than 62000 patterns of the mass spectrum. During matching with NIST library the match factor greater than 700 was considered only for better and pure results. The results revealed that different solvent extracted samples analyzed through GC-MS contained appreciable quantities of different bioactive molecules including antibiotics, fatty acids and protein which have important pharmacological significance.

Antinociceptive, antimicrobial potential and phytochemical screening of different solvent extracted samples from the stem of Acer pentapomicum.

The present study investigates the antinociceptive, antimicrobial activity and phytochemical assessment of samples from A. pentapomicum. Different microbes were tested using disc diffusion assay at three concentrations (1, 2 and 3 mg/disc). Antinociceptive activity was determined by acetic acid induced and hot plate methods. The tested plant extracts revealed significant antinociceptive activity at dose dependent manner when measured by acetic acid induced method. The growth of all the tested microbes was inhibited by methanol, butanol, and ethyl acetate extracted samples at all concentrations. Chloroform, n-hexane and aqueous extracts inhibited the growth of the different microbes at high concentration. The most susceptible microbe was P. aeruginosa measuring 94% and 79% zone of inhibition by butanol and ethyl acetate extracts at 3mg /disc. C. fruendii, however, was the most resistant bacterium followed by S. aureus. The presence of alkaloids, proteins, amino acids, carbohydrates, flavonoids, tannins, saponins, and fats were confirmed in phytochemical screening of different extracts.

Green synthesis and characterization of silver nano-particles from pharmaceutically important Periploca hydaspidis and their biological activity.

The present study describes the synthesis, characterization of nano-particles from Periploca hyaspidis and their in vitro biological activity. The synthesis of AgNPs was confirmed by UV-Vis spectrophotometer and structure by atomic force microscope. The crystallite size and different functional groups was determined by X-ray diffraction and Fourier transformed infrared spectroscopy. Anti-microbial and anti-oxidant activity was carried out by disc diffusion and DPPH radical scavenging protocol respectively. Silver nano-particles (AgNPs) were synthesized by mixing 1mM AgNO3 solutions with plant boiled extract in 1:9. The color change from yellow to dark brown indicated the synthesis of the nano-particles. The AgNPs were more stable at 25°C to 45°C, 1mM concentration of the salt and neutral to slightly basic pH. The results revealed that aromatic amines were responsible for the synthesis of AgNPs. The crystallite size was 7.50 nm, cubic and in nanorgime. AgNPs showed good anti-oxidant activity and was effective against K. pnemoniae, E. coli, X. compestris, C. albicans and P. chrysogenum.

Phytochemical analysis, phytotoxic and insecticidal activities of medicinally important Periploca hydaspidis.

The present study investigates the phyto-chemical analysis, phytotoxic activity and insecticidal activity of the stem and root tissues of medicinally important Periploca hydaspidis. All the extracted samples exhibited the phytotoxic activity. The data confirmed that water was the best solvent to extract the phytotoxic compounds from stem and root tissues. Roots extracted with different solvents exhibited better phytotoxic potential than samples obtained from the stem tissues. Samples extracted in methanol and water from both stems and roots tissues were active against the subject insects while hexane, ethyl acetate and butanol extracted samples in both parts exhibited no insecticidal activity. Water extracted samples of the stem and root exhibited better insecticidal activity compared with methanol extracted samples. Tribolium castaneum and Rhizopertha dominica were more sensitive in both the cases than Callosobruchus maculates. The plant contained alkaloids, flavonoids, carbohydrate, proteins and saponins.

Phytochemical screening and antibacterial activity of different solvent extracted samples of Arisaema jacquemontii.

The current research was carried out to assess the antibacterial activities and phytochemical analysis of the methanol, n-hexane, ethyl acetate, n-butanol soluble fractions and aqueous extracts of the tubers of Arisaema jacquemontii. All the extracts were tested for their antibacterial potential at 1, 2 and 3 mg disc-1 concentrations against 6 bacterial strains through disc diffusion suseptibility assay. The data suggested that different extracts showed varying degree of growth inhibition against the tested microbes. Statistical analysis revealed that n-hexane and ethyl acetate soluble fractions significantly inhibited the growth of all the bacterial strains at the tested concentrations. Moderate activities were recorded for n-butanol and methanolic extracted samples at different concentrations against all the tested strains of bacteria. P. aeruginosa, S. aureus and X. campestris showed resistance to all the tested concentrations of the aqueous extract. B. subtilis and K. pneumoniae were resistant at 1 and 2 mg disc-1 concentrations of the aqueous extract and 3 mg disc-1 of the same extract reduced the growth of the same bacteria. Phytochemical analysis of the different solvent extracted samples suggested the presence or absence of various metabolites including alkaloids, saponins, tannins, sterols, flavonoids, protein, carbohydrates and fats.

Fractionation of crude extracts from controlled dried and commercially available stem bark of Juglans regia and their antimicrobial effects.

The current research investigates the anti-microbial activities of methanol, ethyl acetate, n-hexane , n-butanol and water extracted samples from controlled dried and commercial bark of walnut (Juglans regia) against five bacterial (Staphylococcus aureus, Bacilus subtilis, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonia and one fungal pathogenic specie (Candida albicans) by discs diffusion susceptibility assay using 0.5 and 1mg disc-1 concentrations. Our results revealed that all the extracts from controlled dried and commercial bark of walnut showed varying degrees of antimicrobial activities. Ethyl acetate fraction from both sources exhibited maximum activity against all tested microbial species followed by n-butanol and crude methanolic extraxt. N-hexane and aqueous extracted samples from controlled dried bark reduced the growth of all studied microbes except Staphylococcus aureus in case of commercial available bark. Aqueous extracted sample showed inhibitory effects against all tested microbes except Candida albicans respectively in case of commercial bark. The most susceptible gram positive bacteria were S. aureus while Bacillus subtilis was the most resistant one. Among Gram negative bacteria, Pseudomonas aeruginosa was the most susceptibility while Klebsella pneumonia showed some resistively. Compared to commercial bark samples, controlled dried bark extract and fractions were found to be more active in reducing the growth of all the tested microbes at both concentrations.